心肌细胞面积与其活力缺乏相关关系

No relation between cardiomyocyte surface area and its viability

目的 :建立培养心肌细胞活力的测定与简便的面积测量方法 ,观测不同培养条件下 ,新生大鼠心肌细胞面积与活力的关系 .方法 :采用一步伊红染色法与NIHImage软件测量心肌细胞面积 .MTT比色法测量心肌细胞的A490nm 值 ,以A490nm/总细胞数之比作为衡量心肌细胞活力的指标 .结果 :心肌细胞面积随培养时间延长而增大 ,随接种密度增加而减小 ,且有血清培养组的面积明显大于无血清培养组 (P <0 .0 1 ) .心肌细胞的A490nm值与A490nm/总细胞数之比随培养时间延长均保持不变 ,当细胞接种密度为 4 .5× 1 0 4/cm2 时 ,A490nm/总细胞数之比达峰值 ,且无血清培养组的A490nm值与A490nm/总细胞数之比皆明显低于有血清组 (P <0 .0 1 ) .结论 :在培养时间与细胞接种密度不同的培养条件下 ,培养新生大鼠心肌细胞面积的改变并未对其活力产生明显影响 。

AIM: To set up a simple and effective method to measure the cardiomyocyte surface area and its viability and to study the relation between cardiac myocyte surface area and its viability. METHODS: The cultured neonatal rat cardiac myocytes were stained by rapid eosine staining and the cell surface area was measured by NIH Image software. The absorbency of the cells at 490 nm ( A 490 nm ) was measured by MTT assay and the cardiac myocyte viability was assessed by the ratio of A 490 nm to total cell numbers. RESULTS: The cardiac myocyte surface area increased in 2-7 d of culture. The surface area of the cells cultured in serum containing medium was bigger than that cultured in serum free medium ( P <0.01). The higher the cell seeding density was, the smaller the cell surface area. The total cardiac myocyte A 490 nm value and A 490 nm /total cell numbers did not change significantly in 3-8 d of culture. But A 490 nm value and A 490 nm /total cell numbers of cardiac myocytes cultured in serum free medium were lower than those in serum containing medium ( P <0.01). There was a positive relation between A 490 nm value and cell seeding density and A 490 nm /total cell numbers reached the highest when cell seeding density was 4.5×10 4/cm 2. CONCLUSION: There is no relation between cardiac myocyte surface area and its viability under different culture conditions.